1. Field of the Invention
The present invention pertains to a biologically active mixture of high purity, high molecular weight straight chained primary aliphatic alcohols (referred to herein as aliphatic alcohols) having enhanced purity that is isolated from a naturally occurring source such as beeswax. More particularly the invention pertains to a mixture of high purity, high molecular weight straight chained primary aliphatic alcohols that is obtained from saponified and/or unsaponified beeswax by liquid extraction wherein the resulting aliphatic alcohols in the mixture contain 20 to 34 carbon atoms.
2. Description of the State of Art
All kinds of waxes, and more especially beeswax, have always been a matter of interest. This has been the case not only because of their industrial applications, but also because of their chemical composition. The amount of beeswax in honey ranges between 0.9% to 1.13%, depending on the methods used to separate the wax from the honey. This wax contains esters, hydrocarbons, free fatty acids, free alcohols and a long list of minor compounds.
The natural mixture of aliphatic alcohols obtained from beeswax has been studied by several authors to learn about its composition and main features. The obtaining of different mixtures of aliphatic alcohols from various waxes has also been reported. (J. A. Lamberton, et al., Australian Journal of Chemistry, 13:261-268 (1959); A. Horn and J. S. Martic, Journal of Science Food and Agriculture, 10:571 (1957); Kreger, (1948); Wimbero, (1904); and Mitsui and Col, (1942)). These studies suggest a method for obtaining aliphatic alcohols based on the homogeneous saponification with alcoholic potassium hydroxide.
Another method reports an extraction of the natural aliphatic alcohol mixture through a high efficiency vacuum. The high vacuum wax distillation for the chemical isolation of mixed derivatives and the extraction of the remaining wax are done using petroleum ether. The solvent is evaporated and the remaining solids are acetylated for further purification through alumina chromatography. Finally, through alkaline hydrolysis, aliphatic alcohols are obtained and then recrystallized in ethanol, showing a melting point range from 62xc2x0 C. to 82xc2x0 C.
Blood-lipid lowering effects of a natural mixture of straight chain aliphatic alcohols have been demonstrated by several authors: (F. Liu, et al., xe2x80x9cActive Constituents Lowering Blood-Lipid in Beeswax,xe2x80x9d Zhongguo Zhong Yao Zhi, 21(9):553-4, 576 (1996)); (H. Sho, et al., xe2x80x9cEffects of Okinawa Sugar Cane Wax and Fatty Alcohols on Serum and Liver Lipids in the Rats,xe2x80x9d J Nutri Vitaminol, 30(6):553-559 (1984)); (S. Kato, K. Hamatani, et al., xe2x80x9cOctacosanol Effects Lipid Metabolism in Rat Fed on a High Fat Diet,xe2x80x9d Br J Nutr, 73(3): 433-441 (1995)); and (Kabiry, et al., xe2x80x9cTissue Distribution of Octacosanol in Liver and Muscle of Rats After Serial Administration,xe2x80x9d Ann Nutr Metab, 39(5):279-284 (1995)); and (I. Gouni-Berthold, et al., xe2x80x9cPolicosanol: Clinical pharmacology and therapeutic significance of a new lipid-lowering agent,xe2x80x9d Am Heart J, 143:356-365 (2002)). Many investigational studies based on clinical studies using the natural mixtures of straight chain aliphatic alcohols have been published.
A procedure for obtaining a natural mixture of aliphatic alcohols from animal and vegetable waxes (a natural sourced wax) is also known in the prior art. This prior art procedure is based on the extraction of alcohol mixtures with fluid extractant in the sub- and super-critical states between 20xc2x0 C. and 100xc2x0 C. Selective extraction can be carried out with this procedure, but when this is applied to beeswax it is only possible to obtain between 10% to 15% of a C-20 to C-34 alcohol mixture.
Another project (S. Inaa, K. Furukama, T. Masui, K. Honda, J. Ogasawara, and G. Tsubikamoto, xe2x80x9cProcess for Recovering Primary Normal Aliphatic Higher Alcoholsxe2x80x9d JP 60-119514 (1996)) proposed a very similar extraction method applied to waxes that is based on fluids (CO2 with ethylene) in sub- and super-critical states.
There are several different commercial dietary supplements, foods and drugs that aid in the lowering of total blood cholesterol (lowering lipid, LDL-cholesterol, and total cholesterol levels) which are considered as effective, safe and well-tolerated but most produce different adverse side effects. Since lipid-lowering therapy must be chronically administered, safety and tolerability are very important for their definitive acceptance.
It has been described that treatment with some lipid-lowering drugs reduces the tendency for platelet hyperaggregation frequently seen in hyperlipidemic patients and experimental data have shown anti-aggregatory effects mediated by these compounds. Nevertheless, only some cholesterol-lowering drugs show this property.
Atherosclerosis is a variable combination of changes of the intima of the arteries consisting of the focal accumulation of lipids, complex carbohydrates, blood and blood products, fibrous tissue and calcium deposits, frequently also associated with medial changes. Thus, atherosclerosis is known as a multifactorial process and includes hyperlipidemia as a risk factor.
Among the factors contributing to atherosclerosis development, platelet aggregation has a very important place. The granule contents released from platelets activate arachidonic acid, which metabolizes into cyclic endoperoxides. These are mainly transformed into thromboxane A2 (TXA2), a strong vasoconstrictor and platelet aggregatory agent. Platelet aggregation can be elicited by numerous compounds, such as collagen, ADP, and epinephrine. Thus, different experimental xe2x80x9cin vivo,xe2x80x9d xe2x80x9cex vivo,xe2x80x9d or xe2x80x9cin vitroxe2x80x9d models that test effectiveness of putative antiplatelet drugs commonly test their effect on platelet aggregation induced by these agents. These tests are also used for testing platelet aggregation in healthy volunteers and in patients with diseases which induce hyperaggregability such as hypercholesterolemia and diabetes.
Collagen-induced platelet aggregation is one of the most frequently used tests. Thus, for example, collagen injected intravenously leads to reversible intravascular platelet aggregation xe2x80x9cin vivoxe2x80x9d and aggregates of platelets enter the vascular microcirculation, subsequently decreasing the count of circulating platelets and simultaneously increasing the plasma malondialdehyde (MDA) concentration. Moreover, in some species the injection of collagen induces mortality produced by thrombosis. In these models, antiplatelet drugs generally prevent the decrease in circulating platelet content and the increase of MDA concentration, as well as collagen induced mortality.
Some drugs showing platelet anti-aggregatory effects are useful for treatment of thrombotic diseases, myocardial infarction, and stroke, but not all show these advantages. On the other hand, there are antithrombotic drugs such as streptokinase and urokinase that mainly act by lytic processes affecting blood coagulation, but not on the platelet aggregation. Since ischemic cardiovascular diseases, stroke and vascular peripheric obstructive pathologies are the main sequence of atherosclerosis, effects of several drugs on these complications are commonly tested. Thus, theoretically a drug showing cholesterol lowering properties that also can prevent these complications by acting on other events involved in these processes must be advantageous for treating these patients. Likewise, reduction of TXA2 levels has been associated not only with antiplatelet and antithrombotic effects, but also with antischemic effects. The pharmacological screening of antischemic drugs commonly includes the evaluation of their effects on brain-induced global ischemia. Thus, the protective effect of different drugs on rat cerebral ischemia has been determined by this type of evaluation for certain non-steroidal anti-inflammatory drugs (NSAID) which inhibit reactions catalyzed by cyclooxygenase, as well as for specific inhibitors of thromboxane synthetase and prostacyclin (PGI2) analogues (M. G. Borzeix and J. Cahn, xe2x80x9cEffects of New Chemically Metabolically Stable Prostacyclin Analogues on Early Consequences of a Transient Cerebral Oligemia in Rats,xe2x80x9d Prostaglandins, 35(5):653-664 (1998)). Other experimental models, such as global ischemia induced experimentally in Mongolian gerbils, are also used frequently.
The method and resulting composition of the present invention begins by initially subjecting a natural product containing aliphatic alcohols present as esters to a homogenous phase saponification step after which the saponified product is recovered, dried, and ground to a particle mesh size of 100-500 microns. Alternatively, the saponification step may be skipped entirely and the unsaponified natural product may be dried and ground to a particle mesh size of 100-200 microns.
Next, the particles of natural product containing the aliphatic alcohols (saponified or unsaponified) are placed into a conventional solid-liquid extractor, and a solvent is introduced and heated in contact with the natural product particles. The resulting solution is hot-filtered to remove the solids. The extract is then cooled to the temperature range of 2xc2x0 C.-10xc2x0 C. causing the aliphatic alcohols to solidify and form a suspension. The suspension is filtered, and the solids (first solids) are recovered and air dried. The solids obtained from this step are sent to a purifier where they are contacted with and dissolved in a second hot organic solvent. This solution is then hot-filtered and cooled. The solids (second solids) which form on cooling are again collected and dried by vacuum. The second solids are contacted with another hot organic solvent which dissolves the solids. This solution is hot-filtered and then chilled to resolidify the solids. These third solids are collected, dried and powdered and are the final product.
The final product contains a mixture of aliphatic alcohols from 20 to 34 carbon atoms comprising 1-eicosanol, 1-docosanol, 1-tetracosanol, 1-hexacosanol, 1-heptacosanol, 1-octacosanol, 1-triacontanol, 1-dotriacontanol and 1-tetratriacontanol having the following quantitative composition:
These solids may be used as is or reformulated for administration to humans and animals to reduce and/or prevent hypercholesterolemic diseases, total cholesterol, LDL-cholesterol, coronary heart disease (heart attacks and strokes), inflammation or immunoregulatory diseases, cardiovascular diseases, and/or neurodegenerative disorders. The daily dosage is established between 1 to 100 mg of aliphatic alcohol per day (preferably 3 to 20 mg) and is intended for ingestion in any type or form of foodstuff, capsule, tablet or liquid form.